Pharmaceutical Compositions And Methods

ABSTRACT

Pharmaceutical compositions and kits including a tyrosine hydroxylase inhibitor; melanin, a melanin promoter, or a combination thereof a p450 3A4 promoter; and a leucine aminopeptidase inhibitor are provided. Also provided are methods of treating cancer in a subject, comprising administering an effective amount of a tyrosine hydroxylase inhibitor, a melanin promoter, a p450 3A4 promoter, and a leucine aminopeptidase inhibitor to the subject in need thereof. Also provided are methods of reducing cell proliferation in a subject comprising administering an effective amount of a tyrosine hydroxylase inhibitor, a melanin promoter, a p450 3A4 promoter, and a leucine aminopeptidase inhibitor to the subject in need thereof.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application is a continuation of U.S. patent application Ser. No.14/750,877, filed Jun. 25, 2015, which is a continuation of U.S. patentapplication Ser. No. 13/742,865, filed Jan. 16, 2013, which is acontinuation-in-part of U.S. patent application Ser. No. 13/371,076,filed on Feb. 10, 2012, now U.S. Pat. No. 8,481,498, which claimspriority to U.S. Provisional Application No. 61/587,420, filed on Jan.17, 2012. This application also claims priority to U.S. ProvisionalApplication No. 61/702,994, filed on Sep. 19, 2012. All of theapplications mentioned in this paragraph are incorporated herein byreference in their entirety.

TECHNICAL FIELD

The present inventions relate generally to compositions, kits andmethods for the reduction of cellular proliferation as, for example, inthe treatment of cancer.

BACKGROUND

According to the U.S. National Cancer Institute's SurveillanceEpidemiology and End Results (SEER) database for the year 2008, the mostrecent year for which incidence data are available, 11,958,000 Americanshave invasive cancers. Cancer is the second most common cause of deathin the United States, behind only heart disease, and accounts for one infour deaths. It has been estimated that approximately 1600 Americans dieof cancer each day. In addition to the medical, emotional andpsychological costs of cancer, cancer has significant financial costs toboth the individual and society. It is estimated by the NationalInstitutes of Health that the overall costs of cancer in 2010 was $263.8billion. In addition, it is estimated that another $140.1 billion islost in productivity due to premature death.

Cancer treatments today include surgery, hormone therapy, radiation,chemotherapy, immunotherapy, targeted therapy, and combinations thereof.Surgical removal of cancer has advanced significantly; however, thereremains a high chance of recurrence of the disease. Hormone therapyusing drugs such as aromatase inhibitors and luteinizinghormone-releasing hormone analogs and inhibitors has been relativelyeffective in treating prostate and breast cancers. Radiation and therelated techniques of conformal proton beam radiation therapy,stereotactic radiosurgery, stereotactic radiation therapy,intraoperative radiation therapy, chemical modifiers, and radiosensitizers are effective at killing cancerous cells, but can also killand alter surrounding normal tissue. Chemotherapy drugs such asaminopterin, cisplatin, methotrexate, doxorubicin, daunorubicin andothers alone and in combinations are effective at killing cancer cells,often by altering the DNA replication process. Biological responsemodifier (BRM) therapy, biologic therapy, biotherapy, or immunotherapyalter cancer cell growth or influence the natural immune response, andinvolve administering biologic agents to a patient such as aninterferons, interleukins, and other cytokines and antibodies such asrituximab and trastuzumab and even cancer vaccines such as Sipuleucel-T.

Recently, new targeted therapies have been developed to fight cancer.These targeted therapies differ from chemotherapy because chemotherapyworks by killing both cancerous and normal cells, with greater effectson the cancerous cells. Targeted therapies work by influencing theprocesses that control growth, division, and the spread of cancer cellsand signals that cause cancer cells to die naturally. One type oftargeted therapy includes growth signal inhibitors such as trastuzumab,gefitinib, imatinib, centuximab, dasatinib and nilotinib. Another typeof targeted therapy includes angiogenesis inhibitors such as bevacizumabthat inhibit cancers from increasing surrounding vasculature and bloodsupply. A final type of targeted therapy includes apoptosis-inducingdrugs that are able to induce direct cancer cell death.

Although all of these treatments have been effective to one degree oranother, they all have drawbacks and limitations. In addition to many ofthe treatments being expensive, they also are often too imprecise or thecancers are able to adapt to them and become resistant.

Thus, there is a great need for additional cancer treatments. Inparticular, there is a need for treatments for cancers that have becomeresistant to other forms of treatment.

SUMMARY

The present invention provides compositions, combination therapies,kits, and methods for reducing undue cellular proliferation, includingthat associated with the treatment of cancer. In certain embodiments,the invention provides pharmaceutical compositions comprising at leastone tyrosine hydroxylase inhibitor; at least one of melanin, a melaninpromoter, or a combination thereof; at least one p450 3A4 promoter; atleast one leucine aminopeptidase inhibitor; and, optionally, at leastone growth hormone inhibitor. In other embodiments, the inventionprovides kits that comprise these components together with suitablepackaging. Also provided are methods of reducing cellular proliferationand/or methods of treating cancer comprising administering an effectiveamount of at least one tyrosine hydroxylase inhibitor; at least one ofmelanin, a melanin promoter, or a combination thereof at least one p4503A4 promoter; at least one leucine aminopeptidase inhibitor; and,optionally, at least one growth hormone inhibitor to the subject in needthereof.

DETAILED DESCRIPTION OF ILLUSTRATIVE EMBODIMENTS

The present subject matter may be understood more readily by referenceto the following detailed description which forms a part of thisdisclosure. It is to be understood that this invention is not limited tothe specific products, methods, conditions or parameters describedand/or shown herein, and that the terminology used herein is for thepurpose of describing particular embodiments by way of example only andis not intended to be limiting of the claimed invention.

Unless otherwise defined herein, scientific and technical terms used inconnection with the present application shall have the meanings that arecommonly understood by those of ordinary skill in the art. Further,unless otherwise required by context, singular terms shall includepluralities and plural terms shall include the singular.

As employed above and throughout the disclosure, the following terms andabbreviations, unless otherwise indicated, shall be understood to havethe following meanings.

In the present disclosure the singular forms “a,” “an,” and “the”include the plural reference, and reference to a particular numericalvalue includes at least that particular value, unless the contextclearly indicates otherwise. Thus, for example, a reference to “acompound” is a reference to one or more of such compounds andequivalents thereof known to those skilled in the art, and so forth. Theterm “plurality”, as used herein, means more than one. When a range ofvalues is expressed, another embodiment includes from the one particularand/or to the other particular value. Similarly, when values areexpressed as approximations, by use of the antecedent “about,” it isunderstood that the particular value forms another embodiment. Allranges are inclusive and combinable.

As used herein, the terms “component,” “composition,” “composition ofcompounds,” “compound,” “drug,” “pharmacologically active agent,”“active agent,” “therapeutic,” “therapy,” “treatment,” or “medicament”are used interchangeably herein to refer to a compound or compounds orcomposition of matter which, when administered to a subject (human oranimal) induces a desired pharmacological and/or physiologic effect bylocal and/or systemic action.

As used herein, the terms “treatment” or “therapy” (as well as differentforms thereof) include preventative (e.g., prophylactic), curative orpalliative treatment. As used herein, the term “treating” includesalleviating or reducing at least one adverse or negative effect orsymptom of a condition, disease or disorder. This condition, disease ordisorder can be cancer.

As employed above and throughout the disclosure the term “effectiveamount” refers to an amount effective, at dosages, and for periods oftime necessary, to achieve the desired result with respect to thetreatment of the relevant disorder, condition, or side effect. It willbe appreciated that the effective amount of components of the presentinvention will vary from patient to patient not only with the particularcompound, component or composition selected, the route ofadministration, and the ability of the components to elicit a desiredresult in the individual, but also with factors such as the diseasestate or severity of the condition to be alleviated, hormone levels,age, sex, weight of the individual, the state of being of the patient,and the severity of the pathological condition being treated, concurrentmedication or special diets then being followed by the particularpatient, and other factors which those skilled in the art willrecognize, with the appropriate dosage being at the discretion of theattending physician. Dosage regimes may be adjusted to provide theimproved therapeutic response. An effective amount is also one in whichany toxic or detrimental effects of the components are outweighed by thetherapeutically beneficial effects.

“Pharmaceutically acceptable” refers to those compounds, materials,compositions, and/or dosage forms which are, within the scope of soundmedical judgment, suitable for contact with the tissues of human beingsand animals without excessive toxicity, irritation, allergic response,or other problem complications commensurate with a reasonablebenefit/risk ratio.

Within the present invention, the disclosed compounds may be prepared inthe form of pharmaceutically acceptable salts. “Pharmaceuticallyacceptable salts” refer to derivatives of the disclosed compoundswherein the parent compound is modified by making acid or base saltsthereof. Examples of pharmaceutically acceptable salts include, but arenot limited to, mineral or organic acid salts of basic residues such asamines; alkali or organic salts of acidic residues such as carboxylicacids; and the like. The pharmaceutically acceptable salts include theconventional non-toxic salts or the quaternary ammonium salts of theparent compound formed, for example, from non-toxic inorganic or organicacids. For example, such conventional non-toxic salts include thosederived from inorganic acids such as hydrochloric, hydrobromic,sulfuric, sulfamic, phosphoric, nitric and the like; and the saltsprepared from organic acids such as acetic, propionic, succinic,glycolic, stearic, lactic, malic, tartaric, citric, ascorbic, pamoic,maleic, hydroxymaleic, phenylacetic, glutamic, benzoic, salicylic,sulfanilic, 2-acetoxybenzoic, fumaric, toluenesulfonic, methanesulfonic,ethane disulfonic, oxalic, isethionic, and the like. Thesephysiologically acceptable salts are prepared by methods known in theart, e.g., by dissolving the free amine bases with an excess of the acidin aqueous alcohol, or neutralizing a free carboxylic acid with analkali metal base such as a hydroxide, or with an amine.

Compounds described herein can be prepared in alternate forms. Forexample, many amino-containing compounds can be used or prepared as anacid addition salt. Often such salts improve isolation and handlingproperties of the compound. For example, depending on the reagents,reaction conditions and the like, compounds as described herein can beused or prepared, for example, as their hydrochloride or tosylate salts.Isomorphic crystalline forms, all chiral and racemic forms, N-oxide,hydrates, solvates, and acid salt hydrates, are also contemplated to bewithin the scope of the present invention.

Certain acidic or basic compounds of the present invention may exist aszwitterions. All forms of the compounds, including free acid, free baseand zwitterions, are contemplated to be within the scope of the presentinvention. It is well known in the art that compounds containing bothamino and carboxy groups often exist in equilibrium with theirzwitterionic forms. Thus, any of the compounds described herein thatcontain, for example, both amino and carboxy groups, also includereference to their corresponding zwitterions.

The term “stereoisomers” refers to compounds that have identicalchemical constitution, but differ as regards the arrangement of theatoms or groups in space.

The term “administering” means either directly administering a compoundor composition of the present invention, or administering a prodrug,derivative or analog which will form an equivalent amount of the activecompound or substance within the body.

The terms “subject,” “individual,” and “patient” are usedinterchangeably herein, and refer to an animal, for example a human, towhom treatment, including prophylactic treatment, with thepharmaceutical composition according to the present invention, isprovided. The term “subject” as used herein refers to human andnon-human animals. The terms “non-human animals” and “non-human mammals”are used interchangeably herein and include all vertebrates, e.g.,mammals, such as non-human primates, (particularly higher primates),sheep, dog, rodent, (e.g. mouse or rat), guinea pig, goat, pig, cat,rabbits, cows, horses and non-mammals such as reptiles, amphibians,chickens, and turkeys.

The term “inhibitor” as used herein includes compounds that inhibit theexpression or activity of a protein, polypeptide or enzyme and does notnecessarily mean complete inhibition of expression and/or activity.Rather, the inhibition includes inhibition of the expression and/oractivity of a protein, polypeptide or enzyme to an extent, and for atime, sufficient to produce the desired effect.

The term “promoter” as used herein includes compounds that promote theexpression or activity of a protein, polypeptide or enzyme and does notnecessarily mean complete promotion of expression and/or activity.Rather, the promotion includes promotion of the expression and/oractivity of a protein, polypeptide or enzyme to an extent, and for atime, sufficient to produce the desired effect.

In one embodiment, the present invention provides combination therapiesthat alter the defenses of cancerous cells to oxidative stress. Oneclass of such therapies increases free radical availability to cancerouscells. A representative subclass of such therapies involvesadministration of pharmaceutical compositions comprising a tyrosinehydroxylase inhibitor, melanin or a melanin promoter, a p450 3A4promoter, a leucine aminopeptidase inhibitor, and, optionally, a growthhormone inhibitor. Another subclass involves administration ofpharmaceutical compositions comprising melanin and either a tyrosinehydroxylase inhibitor. Particular components of the pharmaceuticalcomposition are described below.

While not intending to be bound by any particular mechanism ofoperation, tyrosine hydroxylase inhibitors according to the presentinvention function by accumulating in cancer cells and preventing themfrom forming a coating of either lipids or hyaluronan. By preventing thecancer cells from forming a coating of either lipids or hyaluron, thecancer cells are believed to be made more accessible to oxidativestress. Representative tyrosine hydroxylase inhibitors include tyrosinederivatives, which typically are rapidly absorbed by most cancers andinflamed tissues. Representative tyrosine derivatives include one ormore of methyl (2R)-2-amino-3-(2-chloro-4-hydroxyphenyl) propanoate,D-tyrosine ethyl ester hydrochloride, methyl(2R)-2-amino-3-(2,6-dichloro-3,4-dimethoxyphenyl) propanoateH-D-Tyr(TBU)-allyl ester HCl, methyl(2R)-2-amino-3-(3-chloro-4,5-dimethoxyphenyl) propanoate, methyl(2R)-2-amino-3-(2-chloro-3-hydroxy-4-methoxyphenyl) propanoate, methyl(2R)-2-amino-3-(4-[(2-chloro-6-fluorophenyl) methoxy] phenyl)propanoate, methyl (2R)-2-amino-3-(2-chloro-3,4-dimethoxyphenyl)propanoate, methyl (2R)-2-amino-3-(3-chloro-5-fluoro-4-hydroxyphenyl)propanoate, diethyl 2-(acetylamino)-2-(4-[(2-chloro-6-fluorobenzyl) oxy]benzyl malonate, methyl (2R)-2-amino-3-(3-chloro-4-methoxyphenyl)propanoate, methyl (2R)-2-amino-3-(3-chloro-4-hydroxy-5-methoxyphenyl)propanoate, methyl(2R)-2-amino-3-(2,6-dichloro-3-hydroxy-4-methoxyphenyl) propanoate,methyl (2R)-2-amino-3-(3-chloro-4-hydroxyphenyl) propanoate,H-DL-tyr-OME HCl, H-3,5-diiodo-tyr-OME HCl, H-D-3,5-diiodo-tyr-OME HCl,H-D-tyr-OME HCl, D-tyrosine methyl ester hydrochloride, D-tyrosine-omeHCl, methyl D-tyrosinate hydrochloride, H-D-tyr-OMe.HCl, D-tyrosinemethyl ester HCl, H-D-Tyr-OMe-HCl, (2R)-2-amino-3-(4-hydroxyphenyl)propionic acid, (2R)-2-amino-3-(4-hydroxyphenyl) methyl esterhydrochloride, methyl (2R)-2-amino-3-(4-hydroxyphenyl) propanoatehydrochloride, methyl (2R)-2-azanyl-3-(4-hydroxyphenyl) propanoatehydrochloride, 3-chloro-L-tyrosine, 3-nitro-L-tyrosine,3-nitro-L-tyrosine ethyl ester hydrochloride, DL-m-tyrosine,DL-o-tyrosine, Boc-Tyr (3,5-I₂)—OSu, Fmoc-tyr(3-NO₂)—OH, andα-methyl-DL-tyrosine (also known as DL-2-Methyl-3-(4-Hydroxyphenyl)alanine).

The present invention involves the use of at least one of melanin, amelanin promoter, or a combination thereof. Thus, melanin can be used,one or more melanin promoters can be used, and both melanin and one ormore melanin promoters can be used (either in separate dosage forms orin the same dosage form). Melanin promoters according to the presentinvention are chemical compounds that increase the production and/or theactivity of melanin. Increased melanin levels are believed to reduceinflammation (through, for example, suppression of TNF) and exclude thesequestered lymph system. Melanin is a photo catalyst, and can thereforepromote chemical reactions that generate free radicals which, in turn,can become accessible to cancer cells. Representative melanin promotersare methoxsalen and melanotan II.

In some instances, the tyrosine hydroxylase inhibitor is mixed withmelanin in the same dosage form. This association of melanin with thetyrosine hydroxylase inhibitor is believed to facilitate uptake ofmelanin in cancer cells because tyrosine hydroxylase inhibitors are morereadily taken up by such cells. In certain embodiments melanin issolubilized in a solubilizing agent and then mixed with the tyrosinehydroxylase inhibitor by methods known in the art. The solubilizingagent may be removed by standard techniques, such as evaporation,drying, etc. The solubilizing agent may be a non-toxic solubilizingagent, such as hydrogen peroxide or other solubilizing agents commonlyknown in the art. The melanin and/or the pharmaceutical composition maybe further processed to optimize the pharmaceutical composition's effecton cancer cells. In another embodiment the pharmaceutical compositionmay include additional active agents and/or pharmaceutical excipients.

The pharmaceutical compositions of the invention also include a p450 3A4promoter. “Cytochrome p450 3A4” (which can be abbreviated as “p450 3A4”)is a member of the cytochrome p450 superfamily of enzymes, and is amixed-function oxidase that is involved in the metabolism of xenobioticsin the body. It has the widest range of substrates of all of thecytochromes. The function of a p450 3A4 promoter in the pharmaceuticalcompositions of the invention is to increase the expression and/or theactivity of p450 3A4. The increased p450 3A4 expression and/or activityis believed to reduce cortisone and estrogen levels in the patient.Additionally, the increased p450 3A4 expression and/or activity alsoslightly decreases blood pH, which is believed to help to preserve orenhance melanin activity. Representative p450 3A4 promoters are5,5-diphenylhydantoin (sold commercially as, for example, Dilantin),valproic acid, and carbamazepine, which are believed to induceexpression of the p450 3A4 enzyme.

The instant pharmaceutical compositions further include leucineaminopeptidase inhibitors (alternatively known as leucyl aminopeptidaseinhibitors). Leucine aminopeptidases are enzymes that preferentiallycatalyze the hydrolysis of leucine residues at the N-terminus ofpeptides and/or proteins. Inhibiting the expression and/or activity ofleucine aminopeptidases is believed to assist in tumor reabsorption byincreasing cholesterol transport to the liver. Generally, it is believedthat aminopeptidase inhibitors, including aminopeptidase inhibitors,deplete sensitive tumor cells of specific amino acids by preventingprotein recycling, thus generating an antiproliferative effect.Representative leucine aminopeptidase inhibitors areN-[(2S,3R)-3-amino-2-hydroxy-4-phenylbutyryl]-L-leucine, and rapamycin.

The instant pharmaceutical compositions also optionally include a growthhormone inhibitor. Growth hormone (such as, for example, pancreaticgrowth hormone) induces cell replication. Inhibition of the expressionand/or activity of growth hormone is believed to exclude normal cellsfrom rapid replication while allowing cancer cells to continue torapidly replicate and incorporate the tyrosine derivative.Representative growth hormone inhibitors are octreotide, somatostatin,and seglitide.

The pharmaceutical compositions of the invention can further includeD-leucine. D-leucine is a stereoisomer of the naturally occurringL-leucine, the form of leucine incorporated into polypeptides andproteins. D-leucine cannot be incorporated into polypeptides and/orproteins. Along with the leucine aminopeptidase inhibitor, the D-leucineis believed to create a physiological environment that mimics a leucineshortage. Thus, the presence of D-leucine permits the use of lower dosesof leucine aminopeptidase inhibitor in a pharmaceutical composition.

Also provided herein are kits including a combination therapy thatcreates alterations in the defenses of cancerous cells to oxidativestress. An intended suitable embodiment is a kit that includes acombination therapy that increases free radical availability tocancerous cells. Representative kits comprise a tyrosine hydroxylaseinhibitor, melanin and/or a melanin promoter, a p450 3A4 promoter, aleucine aminopeptidase inhibitor and, optionally, a growth hormoneinhibitor of the type described above, together with packaging for same.The kit can include one or more separate containers, dividers orcompartments and, optionally, informational material such asinstructions for administration. For example, each inhibitor or promoter(or the various combinations thereof) can be contained in a bottle,vial, or syringe, and the informational material can be contained in aplastic sleeve or packet or provided in a label. In some embodiments,the kit includes a plurality (e.g., a pack) of individual containers,each containing one or more unit dosage forms of a compound describedherein. For example, the kit can include a plurality of syringes,ampules, foil packets, or blister packs, each containing a single unitdose of a compound described herein or any of the various combinationsthereof. The containers of the kits can be air tight, waterproof (e.g.,impermeable to changes in moisture or evaporation), and/or light-tight.The kit optionally includes a device suitable for administration of thecomposition, e.g., a syringe, inhalant, pipette, forceps, measuredspoon, dropper (e.g., eye dropper), swab (e.g., a cotton swab or woodenswab), or any such delivery device.

Methods of treating cancer in a subject also are provided, as aremethods of reducing undue cellular proliferation. Such methods caninclude administering an effective amount of a combination therapy thatcreates alterations in the defenses of cancerous cells to oxidativestress. Representative methods of treating cancer include administeringan effective amount of a combination therapy that increases free radicalavailability to cancerous cells. Suitable embodiments are methods thatinclude administering an effective amount of the above-noted tyrosinehydroxylase inhibitor, melanin and/or melanin promoter, p450 3A4promoter, leucine aminopeptidase inhibitor and, optionally, growthhormone inhibitor. Other suitable methods include administering aneffective amount of melanin and a tyrosine hydroxylase inhibitor.

Suitable methods include simultaneous or at least contemporaneousadministration of at least two of the tyrosine hydroxylase inhibitor,melanin or a melanin promoter, p450 3A4 promoter, and leucineaminopeptidase inhibitor, at least three of them, or each of them (ineach case, optionally, with a growth hormone inhibitor). It is believedto be desirable that an effective concentration of these moieties be inthe subject's bloodstream at the same time, and any dosing regimen thatachieves this is within the scope of the present invention. The desirednumber of inhibitors and promoters can be provided in a single dosageform or any number of desired dosage forms, including in individualdosage forms. Representative dosage forms include tablets, capsules,caplets, sterile aqueous or organic solutions, reconstitutable powders,elixirs, liquids, colloidal or other types of suspensions, emulsions,beads, beadlets, granules, microparticles, nanoparticles, andcombinations thereof. The amount of composition administered will, ofcourse, be dependent on the subject being treated, the subject's weight,the severity of the condition being treated, the manner ofadministration, and the judgment of the prescribing physician.

Administration of the melanin, promoters, and/or inhibitors can bethrough various routes, including orally, nasally, subcutaneously,intravenously, intramuscularly, transdermally, vaginally, rectally or inany combination thereof. Transdermal administration can be effectedusing, for example, oleic acid, 1-methyl-2-pyrrolidone, ordodecylnonaoxyethylene glycol monoether.

The melanin, promoters and/or inhibitors can be administered during acycle consisting of five to seven days of administering the melanin,promoters and/or inhibitors and one to two days of not administering themelanin, promoters and/or inhibitors. The melanin, promoters and/orinhibitors can be administered over the course of at least six of saidcycles. It can be desirable to administer these components about twohours between meals to facilitate uptake.

The subject to which the instant compositions are administered can be amammal, preferably a human.

In one representative method, 60 mg of the tyrosine derivative isadministered orally and 0.25 mL of a 2 mg/mL suspension of the tyrosinederivative is administered subcutaneously; 10 mg of the methoxsalen isadministered orally and 0.25 mL of a 1 mg/mL suspension of themethoxsalen is administered subcutaneously; 30 mg of the5,5-diphenylhydantoin is administered orally; and 20 mg of theN-[(2S,3R)-3-amino-2-hydroxy-4-phenylbutyryl]-L-leucine is administeredorally.

In certain embodiments, the combination therapy comprises: (i) a dosageform containing melanin (50 mcg) and α-methyl-DL-tyrosine (75 mg); (ii)a dosage form containing 5,5-diphenylhydantoin (15 mg) andα-methyl-DL-tyrosine (75 mg); (iii) a dosage form containing3-amino-2-hydroxy-4-phenylbutyryl]-L-leucine (50 mcg) andα-methyl-DL-tyrosine (75 mg); (iv) a dosage form containing3-amino-2-hydroxy-4-phenylbutyryl]-L-leucine (5 mcg), melanotan II (10mcg), and 5,5-diphenylhydantoin (2 mg); and (v) a dosage form containingα-methyl-DL-tyrosine (5 mg) in NaCl bacteriostatic water. In otherembodiments, the combination therapy comprises: (i) a dosage formcontaining melanin (50 mcg) and α-methyl-DL-tyrosine (75 mg); (ii) adosage form containing 5,5-diphenylhydantoin (15 mg) andα-methyl-DL-tyrosine (75 mg); (iii) a dosage form containing rapamycin(0.2 mg) and α-methyl-DL-tyrosine (75 mg); (iv) a dosage form containingrapamycin (0.15 mcg), melanotan II (10 mcg), and 5,5-diphenylhydantoin(2 mg); and (v) a dosage form containing α-methyl-DL-tyrosine (5 mg) inNaCl bacteriostatic water. Dosages that are two times greater than this,and even four times greater than this, are believed to be both safe andefficacious.

Representative methods include those in which the cancer is non-smallcell lung cancer. In certain embodiments, the non-small cell lung canceris stage IV non-small cell lung cancer. In yet other embodiments, thecancer is ovarian cancer, breast cancer, cervical cancer, pancreaticcancer, stomach cancer, brain cancer, liver cancer, or testicularcancer. The cancer can also be leukemia or lymphoma.

In certain embodiments, one or more of the tyrosine hydroxylaseinhibitor; the melanin promoter; the p450 3A4 promoter; and the leucineaminopeptidase inhibitor is a nucleic acid, protein, antibody orantigen-binding fragment of an antibody.

The present methods can include not only the disclosed administrationstep but also the step of assessing progression of said cancer in saidsubject and/or the extent of cellular proliferation. The assessing stepcan be performed before or after the administering step.

Suitable embodiments can include a pharmaceutical composition comprisinga tyrosine hydroxylase inhibitor, melanin and/or a melanin promoter, ap450 3A4 promoter, and a leucine aminopeptidase inhibitor. Thepharmaceutical composition can further comprise a growth hormoneinhibitor. The growth hormone can be pancreatic growth hormone. Thegrowth hormone inhibitor can be octreotide or somatostatin. The tyrosinehydroxylase inhibitor can be a tyrosine derivative. The tyrosinederivative can be one or more of methyl (2R)-2-amino-3-(2-chloro-4hydroxyphenyl) propanoate, D-tyrosine ethyl ester hydrochloride, methyl(2R)-2-amino-3-(2,6-dichloro-3,4-dimethoxyphenyl) propanoateH-D-Tyr(TBU)-allyl ester HCl, methyl(2R)-2-amino-3-(3-chloro-4,5-dimethoxyphenyl) propanoate, methyl(2R)-2-amino-3-(2-chloro-3-hydroxy-4-methoxyphenyl) propanoate, methyl(2R)-2-amino-3-(4-[(2-chloro-6-fluorophenyl) methoxy] phenyl)propanoate, methyl (2R)-2-amino-3-(2-chloro-3,4-dimethoxyphenyl)propanoate, methyl (2R)-2-amino-3-(3-chloro-5-fluoro-4-hydroxyphenyl)propanoate, diethyl 2-(acetylamino)-2-(4-[(2-chloro-6-fluorobenzyl) oxy]benzyl malonate, methyl (2R)-2-amino-3-(3-chloro-4-methoxyphenyl)propanoate, methyl (2R)-2-amino-3-(3-chloro-4-hydroxy-5-methoxyphenyl)propanoate, methyl(2R)-2-amino-3-(2,6-dichloro-3-hydroxy-4-methoxyphenyl) propanoate,methyl (2R)-2-amino-3-(3-chloro-4-hydroxyphenyl) propanoate,H-DL-tyr-OME HCl, H-3,5-diiodo-tyr-OME HCl, H-D-3,5-diiodo-tyr-OME HCl,H-D-tyr-OME HCl, D-tyrosine methyl ester hydrochloride, D-tyrosine-omeHCl, methyl D-tyrosinate hydrochloride, H-D-tyr-OMe.HCl, D-tyrosinemethyl ester HCl, H-D-Tyr-OMe-HCl, (2R)-2-amino-3-(4-hydroxyphenyl)propionic acid, (2R)-2-amino-3-(4-hydroxyphenyl) methyl esterhydrochloride, methyl (2R)-2-amino-3-(4-hydroxyphenyl) propanoatehydrochloride, methyl (2R)-2-azanyl-3-(4-hydroxyphenyl) propanoatehydrochloride, 3-chloro-L-tyrosine, 3-nitro-L-tyrosine,3-nitro-L-tyrosine ethyl ester hydrochloride, DL-m-tyrosine,DL-o-tyrosine, Boc-Tyr (3,5-I₂)—OSu, Fmoc-tyr(3-NO₂)—OH, andα-methyl-DL-tyrosine. The melanin promoter can be methoxsalen ormelanotan II. The p450 3A4 promoter can be 5,5-diphenylhydantoin. Thep450 3A4 promoter can be valproic acid or carbamazepine. The leucineaminopeptidase inhibitor can beN-[(2S,3R)-3-amino-2-hydroxy-4-phenylbutyryl]-L-leucine or rapamycin.The pharmaceutical compositions of the invention can further compriseD-leucine.

Also provided herein are kits comprising a tyrosine hydroxylaseinhibitor, melanin and/or a melanin promoter, a p450 3A4 promoter, and aleucine aminopeptidase inhibitor, together with packaging for same. Thekit can further comprise a growth hormone inhibitor. The growth hormonecan be pancreatic growth hormone. The growth hormone inhibitor can beoctreotide or somatostatin. The tyrosine hydroxylase inhibitor can be atyrosine derivative. The tyrosine derivative can be one or more ofmethyl (2R)-2-amino-3-(2-chloro-4 hydroxyphenyl) propanoate, D-tyrosineethyl ester hydrochloride, methyl(2R)-2-amino-3-(2,6-dichloro-3,4-dimethoxyphenyl) propanoateH-D-Tyr(TBU)-allyl ester HCl, methyl(2R)-2-amino-3-(3-chloro-4,5-dimethoxyphenyl) propanoate, methyl(2R)-2-amino-3-(2-chloro-3-hydroxy-4-methoxyphenyl) propanoate, methyl(2R)-2-amino-3-(4-[(2-chloro-6-fluorophenyl) methoxy] phenyl)propanoate, methyl (2R)-2-amino-3-(2-chloro-3,4-dimethoxyphenyl)propanoate, methyl (2R)-2-amino-3-(3-chloro-5-fluoro-4-hydroxyphenyl)propanoate, diethyl 2-(acetylamino)-2-(4-[(2-chloro-6-fluorobenzyl) oxy]benzyl malonate, methyl (2R)-2-amino-3-(3-chloro-4-methoxyphenyl)propanoate, methyl (2R)-2-amino-3-(3-chloro-4-hydroxy-5-methoxyphenyl)propanoate, methyl(2R)-2-amino-3-(2,6-dichloro-3-hydroxy-4-methoxyphenyl) propanoate,methyl (2R)-2-amino-3-(3-chloro-4-hydroxyphenyl) propanoate,H-DL-tyr-OME HCl, H-3,5-diiodo-tyr-OME HCl, H-D-3,5-diiodo-tyr-OME HCl,H-D-tyr-OME HCl, D-tyrosine methyl ester hydrochloride, D-tyrosine-omeHCl, methyl D-tyrosinate hydrochloride, H-D-tyr-OMe.HCl, D-tyrosinemethyl ester HCl, H-D-Tyr-OMe-HCl, (2R)-2-amino-3-(4-hydroxyphenyl)propionic acid, (2R)-2-amino-3-(4-hydroxyphenyl) methyl esterhydrochloride, methyl (2R)-2-amino-3-(4-hydroxyphenyl) propanoatehydrochloride methyl (2R)-2-azanyl-3-(4-hydroxyphenyl) propanoatehydrochloride, 3-chloro-L-tyrosine, 3-nitro-L-tyrosine,3-nitro-L-tyrosine ethyl ester hydrochloride, DL-m-tyrosine,DL-o-tyrosine, Boc-Tyr (3,5-I₂)—OSu, Fmoc-tyr(3-NO₂)—OH, andα-methyl-DL-tyrosine. The melanin promoter can be methoxsalen ormelanotan II. The p450 3A4 promoter can be 5,5-diphenylhydantoin,valproic acid or carbamazepine. The leucine aminopeptidase inhibitor canbe N-[(2S,3R)-3-amino-2-hydroxy-4-phenylbutyryl]-L-leucine or rapamycin.The kits of the invention can further comprise D-leucine.

Methods of treating cancer in a subject are also provided comprisingadministering an effective amount of a tyrosine hydroxylase inhibitor,melanin and/or a melanin promoter, a p450 3A4 promoter, and a leucineaminopeptidase inhibitor to the subject in need thereof. In a suitableembodiment, the method of treating cancer can further comprise a growthhormone inhibitor. In certain embodiments, at least two of thecomponents (i.e., melanin, promoters and/or inhibitors) are administeredsimultaneously. In other embodiments, at least three of the componentsare administered simultaneously. Each of the components can beadministered simultaneously. In suitable embodiments, the components areadministered orally, subcutaneously, intravenously, transdermally,vaginally, rectally or in any combination thereof. The transdermaladministration can be done with oleic acid, 1-methyl-2-pyrrolidone, ordodecylnonaoxyethylene glycol monoether. In other embodiments, thecomponents are administered during a cycle consisting of five to sevendays of administering the components and one to two days of notadministering the components. The components can be administered overthe course of at least six of said cycles. The tyrosine hydroxylaseinhibitor can be a tyrosine derivative. The tyrosine derivative can beone or more of methyl (2R)-2-amino-3-(2-chloro-4 hydroxyphenyl)propanoate, D-tyrosine ethyl ester hydrochloride, methyl(2R)-2-amino-3-(2,6-dichloro-3,4-dimethoxyphenyl) propanoateH-D-Tyr(TBU)-allyl ester HCl, methyl(2R)-2-amino-3-(3-chloro-4,5-dimethoxyphenyl) propanoate, methyl(2R)-2-amino-3-(2-chloro-3-hydroxy-4-methoxyphenyl) propanoate, methyl(2R)-2-amino-3-(4-[(2-chloro-6-fluorophenyl) methoxy] phenyl)propanoate, methyl (2R)-2-amino-3-(2-chloro-3,4-dimethoxyphenyl)propanoate, methyl (2R)-2-amino-3-(3-chloro-5-fluoro-4-hydroxyphenyl)propanoate, diethyl 2-(acetylamino)-2-(4-[(2-chloro-6-fluorobenzyl) oxy]benzyl malonate, methyl (2R)-2-amino-3-(3-chloro-4-methoxyphenyl)propanoate, methyl (2R)-2-amino-3-(3-chloro-4-hydroxy-5-methoxyphenyl)propanoate, methyl(2R)-2-amino-3-(2,6-dichloro-3-hydroxy-4-methoxyphenyl) propanoate,methyl (2R)-2-amino-3-(3-chloro-4-hydroxyphenyl) propanoate,H-DL-tyr-OME HCl, H-3,5-diiodo-tyr-OME HCl, H-D-3,5-diiodo-tyr-OME HCl,H-D-tyr-OME HCl, D-tyrosine methyl ester hydrochloride, D-tyrosine-omeHCl, methyl D-tyrosinate hydrochloride, H-D-tyr-OMe.HCl, D-tyrosinemethyl ester HCl, H-D-Tyr-OMe-HCl, (2R)-2-amino-3-(4-hydroxyphenyl)propionic acid, (2R)-2-amino-3-(4-hydroxyphenyl) methyl esterhydrochloride, methyl (2R)-2-amino-3-(4-hydroxyphenyl) propanoatehydrochloride, methyl (2R)-2-azanyl-3-(4-hydroxyphenyl) propanoatehydrochloride, 3-chloro-L-tyrosine, 3-nitro-L-tyrosine,3-nitro-L-tyrosine ethyl ester hydrochloride, DL-m-tyrosine,DL-o-tyrosine, Boc-Tyr (3,5-I₂)—OSu, Fmoc-tyr(3-NO₂)—OH, andα-methyl-DL-tyrosine. In a suitable embodiment of the method, 60 mg ofthe tyrosine derivative is administered orally and 0.25 mL of a 2 mg/mLsuspension of the tyrosine derivative is administered subcutaneously.The melanin promoter can be methoxsalen. In another suitable method, 10mg of the methoxsalen is administered orally and 0.25 mL of a 1 mg/mLsuspension of the methoxsalen is administered subcutaneously. Themelanin promoter can also be melanotan II. The p450 3A4 promoter can be5,5-diphenylhydantoin. In another suitable method, 30 mg of the5,5-diphenylhydantoin is administered orally. The p450 3A4 promoter canalso be valproic acid or carbamazepine. The leucine aminopeptidaseinhibitor can beN-[(2S,3R)-3-amino-2-hydroxy-4-phenylbutyryl]-L-leucine. In anothersuitable method, 20 mg of theN-[(2S,3R)-3-amino-2-hydroxy-4-phenylbutyryl]-L-leucine is administeredorally. The leucine aminopeptidase inhibitor can also be rapamycin. Thegrowth hormone can be pancreatic growth hormone. The growth hormoneinhibitor can be octreotide. The method can further compriseadministering an effective amount of D-leucine. The subject can be amammal and that mammal can be a human. Representative methods includethose in which the cancer is non-small cell lung cancer. In certainembodiments, the non-small cell lung cancer is stage IV non-small celllung cancer. In other embodiments, the cancer is ovarian cancer, breastcancer, cervical cancer, pancreatic cancer, stomach cancer, braincancer, liver cancer, or testicular cancer. In other embodiments, thecancer is leukemia or lymphoma. In other suitable embodiments, thetyrosine hydroxylase inhibitor, the melanin promoter, the p450 3A4promoter, and the leucine aminopeptidase inhibitor is one or more of anucleic acid, protein, antibody or antigen-binding fragment of anantibody. Another suitable embodiment further comprises assessingprogression of said cancer in said subject. The assessing step can beperformed before said administering step or the assessing step can beperformed after said administering step.

Methods of reducing cell proliferation in a subject are also providedcomprising administering an effective amount of a tyrosine hydroxylaseinhibitor; melanin and/or a melanin promoter; a p450 3A4 promoter; and aleucine aminopeptidase inhibitor to the subject in need thereof. In asuitable embodiment, the method of treating cancer can further comprisea growth hormone inhibitor. In certain embodiments, at least two of thecomponents (i.e., melanin, promoters and/or inhibitors) are administeredsimultaneously. In other embodiments, at least three of the componentsare administered simultaneously. Each of the components can beadministered simultaneously. In suitable embodiments, components areadministered orally, subcutaneously, intravenously, transdermally,vaginally, rectally or in any combination thereof. The transdermaladministration can be done with oleic acid, 1-methyl-2-pyrrolidone, ordodecylnonaoxyethylene glycol monoether. In other embodiments, thecomponents are administered during a cycle consisting of five to sevendays of administering the components and one to two days of notadministering the components. The components can be administered overthe course of at least six of said cycles. The tyrosine hydroxylaseinhibitor can be a tyrosine derivative. The tyrosine derivative can beone or more of methyl (2R)-2-amino-3-(2-chloro-4 hydroxyphenyl)propanoate, D-tyrosine ethyl ester hydrochloride, methyl(2R)-2-amino-3-(2,6-dichloro-3,4-dimethoxyphenyl) propanoateH-D-Tyr(TBU)-allyl ester HCl, methyl(2R)-2-amino-3-(3-chloro-4,5-dimethoxyphenyl) propanoate, methyl(2R)-2-amino-3-(2-chloro-3-hydroxy-4-methoxyphenyl) propanoate, methyl(2R)-2-amino-3-(4-[(2-chloro-6-fluorophenyl) methoxy] phenyl)propanoate, methyl (2R)-2-amino-3-(2-chloro-3,4-dimethoxyphenyl)propanoate, methyl (2R)-2-amino-3-(3-chloro-5-fluoro-4-hydroxyphenyl)propanoate, diethyl 2-(acetylamino)-2-(4-[(2-chloro-6-fluorobenzyl) oxy]benzyl malonate, methyl (2R)-2-amino-3-(3-chloro-4-methoxyphenyl)propanoate, methyl (2R)-2-amino-3-(3-chloro-4-hydroxy-5-methoxyphenyl)propanoate, methyl(2R)-2-amino-3-(2,6-dichloro-3-hydroxy-4-methoxyphenyl) propanoate,methyl (2R)-2-amino-3-(3-chloro-4-hydroxyphenyl) propanoate,H-DL-tyr-OME HCl, H-3,5-diiodo-tyr-OME HCl, H-D-3,5-diiodo-tyr-OME HCl,H-D-tyr-OME HCl, D-tyrosine methyl ester hydrochloride, D-tyrosine-omeHCl, methyl D-tyrosinate hydrochloride, H-D-tyr-OMe.HCl, D-tyrosinemethyl ester HCl, H-D-Tyr-OMe-HCl, (2R)-2-amino-3-(4-hydroxyphenyl)propionic acid, (2R)-2-amino-3-(4-hydroxyphenyl) methyl esterhydrochloride, methyl (2R)-2-amino-3-(4-hydroxyphenyl) propanoatehydrochloride, methyl (2R)-2-azanyl-3-(4-hydroxyphenyl) propanoatehydrochloride, 3-chloro-L-tyrosine, 3-nitro-L-tyrosine,3-nitro-L-tyrosine ethyl ester hydrochloride, DL-m-tyrosine,DL-o-tyrosine, Boc-Tyr (3,5-I₂)—OSu, Fmoc-tyr(3-NO₂)—OH, andα-methyl-DL-tyrosine. In a suitable embodiment of the method, 60 mg ofthe tyrosine derivative is administered orally and 0.25 mL of a 2 mg/mLsuspension of the tyrosine derivative is administered subcutaneously.The melanin promoter can be methoxsalen. In another suitable method, 10mg of the methoxsalen is administered orally and 0.25 mL of a 1 mg/mLsuspension of the methoxsalen is administered subcutaneously. Themelanin promoter can also be melanotan II. The p450 3A4 promoter can be5, 5-diphenylhydantoin. In another suitable method, 30 mg of the 5,5-diphenylhydantoin is administered orally. The p450 3A4 promoter canalso be valproic acid or carbamazepine. The leucine aminopeptidaseinhibitor can beN-[(2S,3R)-3-amino-2-hydroxy-4-phenylbutyryl]-L-leucine. In anothersuitable method, 20 mg of theN-[(2S,3R)-3-amino-2-hydroxy-4-phenylbutyryl]-L-leucine is administeredorally. The leucine aminopeptidase inhibitor can also be rapamycin. Thegrowth hormone can be pancreatic growth hormone. The growth hormoneinhibitor can be octreotide. The method can further compriseadministering an effective amount of D-leucine. The subject can be amammal and the mammal can be a human. Representative methods includethose in which the cancer is non-small cell lung cancer. In certainembodiments, the non-small cell lung cancer is stage IV non-small celllung cancer. In other embodiments, the cancer is ovarian cancer, breastcancer, cervical cancer, pancreatic cancer, stomach cancer, braincancer, liver cancer, or testicular cancer. In other embodiments, thecancer is leukemia or lymphoma. In other suitable embodiments, thetyrosine hydroxylase inhibitor, the melanin promoter, the p450 3A4promoter, and the leucine aminopeptidase inhibitor is one or more of anucleic acid, protein, antibody or antigen-binding fragment of anantibody. Another suitable embodiment further comprises assessingprogression of said cancer in said subject. The assessing step can beperformed before said administering step or the assessing step can beperformed after said administering step.

The following examples of specific embodiments for carrying out thepresent invention are offered for illustrative purposes only, and arenot intended to limit the scope of the present invention in any way.

Representative methods of administration of the pharmaceuticalcompositions and combination therapies also are provided. Variousembodiments of the present invention further relate to methods ofadministering a pharmaceutical composition or combination therapy to ahuman patient for the treatment of cancer. The methods may compriseadministering a pharmaceutical composition or combination therapy bygenerally accepted routes of administration (e.g., oral, subcutaneous,parenteral, inhalation, topical, etc.). In some instances, apharmaceutical composition or combination therapy may be administeredorally and/or subcutaneously. In some instances, a pharmaceuticalcomposition or combination therapy may be administered to human patientsbetween meals.

In certain embodiments of the present invention, a pharmaceuticalcomposition or combination therapy may be administered to a humanpatient for 5 days per week for a period of 6 weeks, creating one cycleof 30 days of treatment. Depending on the outcome after 6 weeks or onecycle of treatment, additional cycles of the pharmaceutical compositionor combination therapy may be administered.

The present invention also provides:

-   -   pharmaceutical compositions comprising a tyrosine hydroxylase        inhibitor; and melanin, a melanin promoter, or a combination        thereof (preferably melanin);    -   pharmaceutical compositions comprising a tyrosine hydroxylase        inhibitor and a p450 3A4 promoter;    -   pharmaceutical compositions comprising a tyrosine hydroxylase        inhibitor and a leucine aminopeptidase inhibitor; and    -   pharmaceutical compositions comprising melanin, a melanin        promoter, or a combination thereof (preferably melanotan II); a        p450 3A4 promoter; and a leucine aminopeptidase inhibitor.

The tyrosine hydroxylase inhibitor in such compositions preferably isα-methyl-DL-tyrosine, the p450 3A4 promoter preferably is5,5-diphenylhydantoin, and the leucine aminopeptidase inhibitorpreferably is N-[(2S,3R)-3-amino-2-hydroxy-4-phenylbutyryl]-L-leucine.The invention also provides kits comprising each of these pharmaceuticalcompositions, along with a pharmaceutical composition comprising atyrosine hydroxylase inhibitor; as well as methods comprisingadministering each of the pharmaceutical compositions to a patient alongwith a pharmaceutical composition comprising a tyrosine hydroxylaseinhibitor, preferably such that they are administered to the patientwithin a 24 hour period.

Example 1

A clinical study was performed to evaluate the effectiveness, safety,acceptability, and tolerability of a combination therapy in accordancewith embodiments of the present invention as a treatment for metastaticcancer.

The combination therapy comprised the following:

-   -   (a) capsule containing melanin (50 mcg) and α-methyl-DL-tyrosine        (75 mg), administered orally;    -   (b) capsule containing 5,5-diphenylhydantoin (15 mg) and        α-methyl-DL-tyrosine (75 mg), administered orally;    -   (c) capsule containing 5,5-diphenylhydantoin (15 mg) and        α-methyl-DL-tyrosine (75 mg), administered orally;    -   (d) capsule containing rapamycin (0.2 mg) and        α-methyl-DL-tyrosine (75 mg), administered orally;    -   (e) suspension containing rapamycin (0.15 mcg), melanotan II (10        mcg), and 5,5-diphenylhydantoin (2 mg), administered        subcutaneously; and    -   (f) suspension containing α-methyl-DL-tyrosine (5 mg) in NaCl        bacteriostatic water, administered subcutaneously.

Each patient was administered the combination therapy five days per weekfor six weeks. More than 200 patients were screened. The criteriaincluded patients with any metastatic cancer. Thirty patients meetingthe criteria were accepted and participated in the study.

Treatment of Metastatic Breast Cancer

Fourteen patients in the study suffered from metastatic breast cancer.Patient information and results are as follows in Table 1:

TABLE 1 Patient Characteristics Average age 55 (40-70 years old) Female14/14  Caucasian 13/14  Prior to Study Declined routine treatment priorto study enrollment 4/14 Used all available treatment and wereconsidered 10/14  incurable Treatment Results 1-3 point improvement inECOG rating 11/14  1-5 point improvement in EORTC rating (scale 1-7)10/14  Weight Gained weight 4/14 (1-5 lbs) Remained the same weight 6/14Lost weight 4/14 (1-2 lbs) Pain Reduction in pain level (scale of 1-10)8/14 (1-9)   Entered study with no pain and maintained the same level6/14 Entered study on pain medication 6/14 No longer needed painmedication at the end of the cycle 5/6  Evidence of Cancer Disease freewith normal physical exam, review of 3/14 systems, and imagingSignificant reduction in quantity and/or size of the largest 5/14 tumorReduction in quantity and/or size of the largest 2/14 tumor Noprogression of the cancer 4/14 Survival Alive 14/14  33-37 wks 4/1427-29 wks 5/14 12-19 wks 5/14 Current Status Went home 3/14 Continuedtreatment 11/14 One side effect of the therapy was hyperpigmentation in all of thepatients. Overall, all of the patients tolerated the combination therapyand no adverse events were reported.

Over 200 cancer patients were screened in a clinical trial. Thirty (30)subjects meeting the study criteria consented. The average patient agewas 56 years old with a range of 30 years old to 70 years old. Thepatients in the study were administered a treatment regimen thatincluded a tyrosine hydroxylase inhibitor (i.e., α-methyl-DL tyrosine),a melanin promoter (i.e., melanotan II), a p450 3A4 promoter (i.e., 5,5-diphenylhydantoin), and a leucine aminopeptidase inhibitor (i.e.,rapamycin). These compounds were administered on each of five days perweek for a period of six weeks, with one or two days off between weeklycycles.

After six weeks of treatment, 12 of the 30 patients (40%) maintained thesame rating under the Eastern Cooperative Oncology Group (ECOG) 0-5scale (see Oken, et al., Toxicity And Response Criteria Of The EasternCooperative Oncology Group, Am. J. Clin. Oncol., 5:649-655, 1982).Fourteen (14) of 30 (46%) had 1-3 point improvement in their ECOGrating.

Fourteen (14) of the 30 patients (46%) maintained the same rating underthe European Organisation for the Research and Treatment of CancerQuality of Life Questionnaire Core (EORTC) 1-7 scale (see, e.g.,Bergman, et al., The EORTC QLQ-LC13: a modular supplement to the EORTCCore Quality of Life Questionnaire (QLQ-C30) for use in lung cancerclinical trials, EORTC Study Group on Quality of Life, Eur. J. Cancer,1994. 30A(5): p. 635-42). Sixteen (16) of 30 (54%) had 1-5 pointimprovement in their EORTC rating.

Eleven (11) of the 30 patients gained weight of 1 to 9 pounds, 17 of 30stayed the same weight, and 2 of 30 lost 1 to 2 pounds.

Thirteen (13) of 30 (43%) had a reduction in pain levels. Seventeen (17)of 30 (57%) entered with minimal pain and maintained the same level.Nine (9) of 30 (30%) entered the study on pain medication and 8 of thosenine (89%) no longer needed pain medication at the end of the cycle.

No disease was detected in four (4) of 30 (13%) with normal physicalexam, review of systems, and imaging. Eight (8) of 30 (27%) hadsignificant reduction in quantity of tumors and/or size of the largesttumor. Eight (8) of 30 (27%) exhibited reduction in quantity of tumorsand/or size of the largest tumor. Ten (10) of 30 (33%) showed noprogression of disease.

Twenty nine (29) of the 30 patients were alive with median survival of22 weeks. Thirteen (13) of 30 (43%) were released and went home.Seventeen (17) of 30 (57%) continue with the treatment. All of thesubjects developed hyperpigmentation.

Overall, the above-noted treatment was well tolerated by the subjects,with no adverse events related to the treatment, and responses have beendocumented to the treatment 100%.

Example 2

In one aspect, the present invention provides methods of inducingmelanin production in vivo with one or both of methoxsalen and melanotanand/or through administration of melanin. Without intending to be boundby any particular theory of operation, melanin is believed to bebeneficial because of its photocatalytic nature and its ability toconvert various wavelengths of ambient or induced electromagneticradiation into electrical energy, thus potentiating desirable reactionsor dislocations. In some patients, either because of genetic variation,infirmity, necessity of expedited availability, or to realize maximumeffectiveness, it has been determined that melanin preferably iscombined mechanically or chemically with α-methyl-DL-tyrosine prior toadministration.

Melanin as a photocatalyst is believed to have polarity at points in itsphysical mass. It has been determined that small melanin particles mayproduce less electrical energy than larger particles, and a plurality ofmelanin particles tend not to accumulate in polarity-specificformations. An effective method to obtain high yield of electricalenergy from melanin is to form the melanin in large, polarizedparticles. By implication, it is believed that nanoparticles, regardlessof quantity, are not as desirable as larger particles for cancertreatment, and that larger particles have a greater capacity to beaccepted by cancer cell membranes.

In accordance with certain embodiments, melanin is combined withα-methyl-DL-tyrosine in at least three ways.

-   -   1) Melanin, either naturally occurring or synthetic, is        mechanically mixed with compressive force to adhere the melanin,        which is non-water soluble and somewhat malleable, with the        α-methyl-DL-tyrosine. Following the initial combining of these        components, it is desirable to add additional        α-methyl-DL-tyrosine until substantial coverage of the melanin        is achieved.    -   2) Melanin can be solubilized by many methods as described in        U.S. Pat. No. 5,225,435, the contents of which are incorporated        herein by reference. One preferred method involves mixing        melanin with distilled water and hydrogen peroxide to achieve a        melanin concentration of at least 5 weight percent, and then        placing the resulting composition in a microwave oven until it        reaches a boiling point. The dissolved melanin that is produced        is used to infuse or saturate through the mass of        α-methyl-DL-tyrosine. The composition is then dried and the dry        powder is used.    -   3) α-Methyl-DL-tyrosine is placed in distilled water with        5-benzyloxy-6-methoxy-indole and sealed for up to a month. The L        portion of the racemic α-methyl-DL-tyrosine is believed to        convert to (DOPA) melanin. The size of the melanin particle can        be controlled by controlling the time of growth period. The        powder produced is then cleaned and dried. The ratio of the        racemic mix is no longer 50/50, but the utility of the        chemically combined ingredients facilitates penetration of the        melanin even with reduced L component and appears to offer        potentially sufficient benefit.

What is claimed:
 1. A method of treating pancreatic cancer in a subjectcomprising administering an effective amount of a tyrosine hydroxylaseinhibitor; a melanin promoter; a p450 3A4 promoter; and a leucineaminopeptidase inhibitor to the subject in need thereof, wherein: themelanin promoter is methoxsalen or melanotan II; the p450 3A4 promoteris 5, 5-diphenylhydantoin, valproic acid, or carbamazepine; and theleucine aminopeptidase inhibitor isN-[(2S,3R)-3-amino-2-hydroxy-4-phenylbutyryl]-L-leucine or rapamycin. 2.The method of claim 1 further comprising administering a growth hormoneinhibitor.
 3. The method of claim 1 wherein at least two of thepromoters and inhibitors are administered simultaneously.
 4. The methodof claim 1 wherein at least three of the promoters and inhibitors areadministered simultaneously.
 5. The method of claim 1 wherein each ofthe promoters and inhibitors is administered simultaneously.
 6. Themethod of claim 1 wherein the promoters and inhibitors are administeredorally, subcutaneously, intravenously, transdermally, vaginally,rectally or in any combination thereof.
 7. The method of claim 6 whereinthe transdermal administration is done with oleic acid,1-methyl-2-pyrrolidone, or dodecylnonaoxyethylene glycol monoether. 8.The method of claim 1 wherein the promoters and inhibitors areadministered during a cycle consisting of five to seven days ofadministering the promoters and inhibitors and one to two days of notadministering the promoters and inhibitors.
 9. The method of claim 8wherein the promoters and inhibitors are administered over the course ofat least six of said cycles.
 10. The method of claim 1 wherein thetyrosine hydroxylase inhibitor is a tyrosine derivative.
 11. The methodof claim 10 wherein the tyrosine derivative is one or more of methyl(2R)-2-amino-3-(2-chloro-4 hydroxyphenyl) propanoate, D-tyrosine ethylester hydrochloride, methyl(2R)-2-amino-3-(2,6-dichloro-3,4-dimethoxyphenyl) propanoateH-D-Tyr(TBU)-allyl ester HCl, methyl(2R)-2-amino-3-(3-chloro-4,5-dimethoxyphenyl) propanoate, methyl(2R)-2-amino-3-(2-chloro-3-hydroxy-4-methoxyphenyl) propanoate, methyl(2R)-2-amino-3-(4-[(2-chloro-6-fluorophenyl) methoxy] phenyl)propanoate, methyl (2R)-2-amino-3-(2-chloro-3,4-dimethoxyphenyl)propanoate, methyl (2R)-2-amino-3-(3-chloro-5-fluoro-4-hydroxyphenyl)propanoate, diethyl 2-(acetylamino)-2-(4-[(2-chloro-6-fluorobenzyl) oxy]benzyl malonate, methyl (2R)-2-amino-3-(3-chloro-4-methoxyphenyl)propanoate, methyl (2R)-2-amino-3-(3-chloro-4-hydroxy-5-methoxyphenyl)propanoate, methyl(2R)-2-amino-3-(2,6-dichloro-3-hydroxy-4-methoxyphenyl) propanoate,methyl (2R)-2-amino-3-(3-chloro-4-hydroxyphenyl) propanoate,H-DL-tyr-OMe HCl, H-3,5-diiodo-tyr-OMe HCl, H-D-3,5-diiodo-tyr-OME HCl,H-D-tyr-OMe HCl, D-tyrosine methyl ester hydrochloride, D-tyrosine-omeHCl, methyl D-tyrosinate hydrochloride, HD-tyr-OMe.HCl, D-tyrosinemethyl ester HCl, H-D-Tyr-OMe-HCl, (2R)-2-amino-3-(4-hydroxyphenyl)propionic acid, (2R)-2-amino-3-(4-hydroxyphenyl) methyl esterhydrochloride, methyl (2R)-2-amino-3-(4-hydroxyphenyl) propanoatehydrochloride, methyl (2R)-2-azanyl-3-(4-hydroxyphenyl) propanoatehydrochloride, 3-chloro-L-tyrosine, 3-nitro-L-tyrosine,3-nitro-L-tyrosine ethyl ester hydrochloride, DL-m-tyrosine,DL-o-tyrosine, Boc-Tyr (3,5-I₂)—OSu, Fmoc-tyr(3-NO₂)—OH, andα-methyl-DL-tyrosine.
 12. The method of claim 11 wherein the tyrosinederivative is α-methyl-DL tyrosine.
 13. The method of claim 11 wherein60 mg of the tyrosine derivative is administered orally and 0.25 mL of a2 mg/mL suspension of the tyrosine derivative is administeredsubcutaneously.
 14. The method of claim 1 wherein the melanin promoteris methoxsalen.
 15. The method of claim 14 wherein 10 mg of themethoxsalen is administered orally and 0.25 mL of a 1 mg/mL suspensionof the methoxsalen is administered subcutaneously.
 16. The method ofclaim 1 wherein the melanin promoter is melanotan II.
 17. The method ofclaim 1 wherein the p450 3A4 promoter is 5, 5-diphenylhydantoin.
 18. Themethod of claim 17 wherein 30 mg of the 5, 5-diphenylhydantoin isadministered orally.
 19. The method of claim 1 wherein the p450 3A4promoter is valproic acid or carbamazepine.
 20. The method of claim 1wherein the leucine aminopeptidase inhibitor isN-[(2S,3R)-3-amino-2-hydroxy-4-phenylbutyryl]-L-leucine.
 21. The methodof claim 20 wherein 20 mg of theN-[(2S,3R)-3-amino-2-hydroxy-4-phenylbutyryl]-L-leucine is administeredorally.
 22. The method of claim 1 wherein the leucine aminopeptidaseinhibitor is rapamycin.
 23. The method of claim 20 wherein the p450 3A4promoter is 5,5-diphenylhydantoin.
 24. The method of claim 23 whereinthe melanin promoter is melanotan II.
 25. The method of claim 1 furthercomprising administering an effective amount of D-leucine.
 26. Themethod of claim 24 wherein the tyrosine derivative is α-methyl-DLtyrosine.
 27. The method of claim 1 wherein the melanin promoter ismelanotan II; the p450 3A4 promoter is 5, 5-diphenylhydantoin; and thetyrosine hydroxylase inhibitor is α-methyl-DL tyrosine.
 28. The methodof claim 27 wherein the leucine aminopeptidase inhibitor isN-[(2S,3R)-3-amino-2-hydroxy-4-phenylbutyryl]-L-leucine.
 29. The methodof claim 27 wherein the leucine aminopeptidase inhibitor is rapamycin.30. The method of claim 1 further comprising assessing progression ofthe pancreatic cancer in said subject.
 31. The method of claim 2 furthercomprising assessing progression of the pancreatic cancer in saidsubject.